Overview

  • Product nameAnti-Calreticulin antibodySee all Calreticulin primary antibodies ...
  • Description
    Rabbit polyclonal to Calreticulin
  • SpecificityThis antibody is specific for calreticulin.
  • Tested applicationsICC/IF, IHC-P, WB, IP, ICC more details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Hamster, Cow, Human, Zebrafish, African Green Monkey
  • Immunogen

    Fusion protein, containing amino acids 154-347 of Mouse Calreticulin.

  • Positive control
    • IC: COS7 and HCT29 cells WB: H4IIE cell lysate, NBL-1 bovine renal epithelial cell lysate

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.05% Sodium Azide
    Constituents: PBS
  • PurityWhole antiserum
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab4 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ICC/IF 1/1000.
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Incubate primary antibody for 1 hour.
WB 1/1000. Detects a band of approximately 55-60 kDa (predicted molecular weight: 48 kDa).
IP 1/300.
ICC 1/250.

Target

  • FunctionMolecular calcium-binding chaperone promoting folding, oligomeric assembly and quality control in the ER via the calreticulin/calnexin cycle. This lectin interacts transiently with almost all of the monoglucosylated glycoproteins that are synthesized in the ER. Interacts with the DNA-binding domain of NR3C1 and mediates its nuclear export.
  • Sequence similaritiesBelongs to the calreticulin family.
  • DomainCan be divided into a N-terminal globular domain, a proline-rich P-domain forming an elongated arm-like structure and a C-terminal acidic domain. The P-domain binds one molecule of calcium with high affinity, whereas the acidic C-domain binds multiple calcium ions with low affinity.
    The interaction with glycans occurs through a binding site in the globular lectin domain.
    The zinc binding sites are localized to the N-domain.
    Associates with PDIA3 through the tip of the extended arm formed by the P-domain.
  • Cellular localizationEndoplasmic reticulum lumen. Cytoplasm > cytosol. Secreted > extracellular space > extracellular matrix. Cell surface. Also found in cell surface (T cells), cytosol and extracellular matrix. Associated with the lytic granules in the cytolytic T-lymphocytes.
  • Target information above from: UniProt accession P27797 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • Autoantigen RO antibody
    • Autoantigen RO antibody
    • CALR antibody
    • CALR antibody
    • CALR protein antibody
    • CALR_HUMAN antibody
    • Calregulin antibody
    • Calreticulin antibody
    • cC1qR antibody
    • CRP55 antibody
    • CRT antibody
    • CRTC antibody
    • Endoplasmic reticulum resident protein 60 antibody
    • ERp60 antibody
    • FLJ26680 antibody
    • grp60 antibody
    • HACBP antibody
    • RO antibody
    • Sicca syndrome antigen A antibody
    • SSA antibody
    see all

Anti-Calreticulin antibody images

  • All lanes : Anti-Calreticulin antibody (ab4) at 1/1000 dilution

    Lane 1 : Chicken lymphoid cell line lysate
    Lane 2 : Chicken lymphoid cell line lysate

    Lysates/proteins at 1000000 cells per lane.

    Secondary
    Polyclonal HRP-conjugate Goat anti-rabbit IgG at 1/2000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 48 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • IF staining was performed on HCT15 colon cancer cells using a 1:50 dilution of the primary antibody followed by hybridisation with a 1:100 dilution of the secondary anti-rabbit antibody Alexa Fluor 488 (molecular Probes) with green fluorescence. Picture from review by Karin Birkenkamp-Demtroeder submitted 20 July 2004
  • ICC/IF image of ab4 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • ab4 staining calreticulin in Human placenta.
    Left panel: with primary antibody diluted 1:4000. Right panel: isotype control.
    Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

References for Anti-Calreticulin antibody (ab4)

This product has been referenced in:
  • Toptan T  et al. Complex alternative cytoplasmic protein isoforms of the Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen 1 generated through noncanonical translation initiation. J Virol 87:2744-55 (2013). Read more (PubMed: 23255808) »
  • Li Q  et al. Estradiol accelerates the effects of fluoxetine on serotonin 1A receptor signaling. Psychoneuroendocrinology 38:1145-57 (2013). WB ; Rat . Read more (PubMed: 23219224) »

See all 29 Publications for this product

Product Wall

Application Western blot
Loading amount 1e+006 cells
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Chicken Cell lysate - other (lymphoid cell line)
Specification lymphoid cell line
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
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Verified customer

Submitted Jul 01 2013

Muchas gracias por tu respuesta y tu comprensión.

Acabo de emitir una nota de abono, con código XXXXXX. Dicha nota de abono debe utilizarse siguiendo uno de los siguientes pasos:

Canjear usando la factura original si t...

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Thank you for contacting Abcam. We do not stock any sample sizes of our antibodies, but we do cover our antibodies under our Abpromise. In this case ab116648, anti-Estradiol antibody, is guaranteed to work in RIA applications. ab116648 has a total of 1...

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Application Western blot
Sample Human Cell lysate - whole cell (HeLa, cervical carcer)
Loading amount 20 µg
Specification HeLa, cervical carcer
Gel Running Conditions Reduced Denaturing (12 %)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Apr 27 2010

Application Western blot
Sample Mouse Cell lysate - whole cell (macrophages)
Loading amount 15 µg
Specification macrophages
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Feb 11 2010

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Neuroblastoma)
Specification Neuroblastoma
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1
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Submitted Feb 12 2007

Thank you for your e-mail and for replying to my further questions. Temperature and incubation period are two crucial factors in the staining procedure. If you incubate the cells with the primary antibody at lower temperature i.e. at 4oC, it is imp...

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Thank you for getting back to me with details of your technical approach. Further to our telephone conversation I have searched the literature and I have determined that Calreticulin antibody - ER Marker (ab4) was used in the following publication;...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa, cerival carcinoma)
Specification HeLa, cerival carcinoma
Fixative Formaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
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Submitted Apr 18 2006

We are not familiar with the method according to Towbin. 1. Can you describe this method in detail? 2. Did the antibody arrive in good condition? at a cold temperature? 3. Did you run any negative controls? If the recombinant protein did not work...

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